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GenomONE™ Series Q&A

Q: Characteristics of HVJ-E
A:
HVJ-E is a non-viral transfection reagent. Since genomic RNA of which has been completely inactivated, it has neither infective nor proliferative potentials in humans or animals.
The molecule to be transferred (DNA, protein, antisense oligonucleotide, siRNA, etc.) is incorporated in HVJ-E particles to yield an HVJ-E vector, which is then introduced into the target cell or tissue, making use of the sialic acid-binding activity of HN protein and membrane-fusion activity of F (fusion) protein existing in HVJ-E.
Q: Method in inactivation of HVJ
A:
Although HVJ-E uses HVJ (Sendai virus/ Murine parainfluenza virus 1) as a raw material, the genomic RNA of HVJ has been completely inactivated by drug treatment*. The HVJ-E will not proliferate or exhibit pathogenic effects in humans or animals.
* Reference:
Prior, P. et al.: BioPharm, 22-33 (Oct. 1996)
Kaneda, Y. et al.: Advances in Genetics, Vol. 53, pp308-332 (2005).
Q: Assessment and confirmation of viral inactivation
A:
Inactivation of HVJ has been confirmed for each lot by the viral proliferative potential rule-out test, using cultured cells and growing chicken eggs.
Q: Bio-safety level for laboratory use
A:
GenomONE series can be used safely in ordinary laboratories. Although HVJ-E uses HVJ (Sendai virus/Murine parainfluenza virus 1) as a raw material, the genomic RNA of HVJ has been completely inactivated by drug treatment. HVJ-E will not proliferate or exhibit pathogenic effects in humans or animals. However, when using this product for recombinant DNA experiments, rules for recombinant DNA experiments (stipulated in relevant statutes in the country of use or set forth by the safety committee of the facility concerned) must be followed, and experiments should only be carried out in laboratories properly equipped with facilities appropriate for recombinant DNA experiments.
Q: Limit of size of molecules that can be incorporated into HVJ-E
A:
Plasmid DNAs of 10 to 15 kbp in size are successfully incorporated into HVJ-E particles and introduced to target cells. Fluorescence-labeled proteins with their molecular weight of 7kDa (insulin) to 150kDa (rabbit IgG) are successfully incorporated into HVJ-E particles. Synthetic compounds (molecular weight >1,000) can also be incorporated into HVJ-E. Limit of the molecular size to be incorporated into the HVJ-E has yet to be clearly demonstrated.
Q: Precaution for in vivo transfection (route of administration)
A:

Because HVJ-E can be adsorbed onto tissue in vivo, especially blood cells through the HN envelope protein, the efficiency of transfection in vivo through intravenous (systemic) administration is usually very low. It is advisable to select a route of administration involving less exposure to blood or to perform perfusion of the animal prior to administration. However, transfections through intravenous injection into mouse has been reported in the following articles.

  1. 1)
    Matsuda N., et al.; Nuclear factor κ-B decoy oligodeoxynucleotides prevent acute lung injury in mice with cecal ligation and puncture-induced sepsis.
    Mol. Pharmacol., 67 (4), 1018-1025 (2005).
  2. 2)
    Takeno M. al. : Th1-dominant shift of T cell cytokine production, and subsequent reduction of serum immunoglobulin E response by administration in vivo of plasmid expressing Txk/Rlk, a member of Tec family tyrosine kinase, in a mouse model.
    Clin. Exp. Allergy, 34, 965-970 (2004).
  3. 3)
    Kaneda Y. et al.; Hemagglutinating virus of Japan (HVJ) envelope vector as a versatile gene delivery system.
    Mol.Ther., 6 (2), 219-226 (2002).

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